Graduate Program in Health Science, State University of Maringá, Colombo Avenue, 5790, Zip Code 87020-900, Maringá, Paraná, Brazil.
Department of Pathological Sciences, Laboratory of Immunoparasitology of Neglected Diseases and Cancer - LIDNC, Center of Biological Sciences, State University of Londrina, Rodovia Celso Garcia Cid Campus, Zip Code 86057-970, Postal box 10.011, Londrina, Paraná, Brazil.
Parasitology. 2021 Oct;148(12):1447-1457. doi: 10.1017/S0031182021001086. Epub 2021 Jun 30.
Toxoplasma gondii is the causative agent of toxoplasmosis, and an important problem of public health. The current treatment for toxoplasmosis is the combination of pyrimethamine and sulphadiazine, which do not act in the chronic phase of toxoplasmosis and have several side-effects. This study evaluated the anti-T. gondii activity and potential mechanism of Moringa oleifera seeds’ aqueous extract in vitro. The concentration of M. oleifera extract in HeLa cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability assays. The presence of T. gondii was assessed by quantitative polymerase chain reaction and toluidine blue staining. Pyrimethamine and sulphadiazine were used as drug controls. Modifications in T. gondii morphology and ultrastructure were observed by electron microscopy. In vitro, the M. oleifera extract had no toxic effect on HeLa cells at concentrations below 50 μg mL−1. Moringa oleifera extract inhibits T. gondii invasion and intracellular proliferation with similar results for sulphadiazine + pyrimethamine, and also shows cellular nitric oxide production at a concentration of 30 μg mL−1. Electron microscopy analyses indicated structural and ultrastructural modifications in tachyzoites after treatment. We also observed an increase in reactive oxygen species production and a loss of mitochondrial membrane integrity. Nile Red staining assays demonstrated a lipid accumulation. Annexin V–fluorescein isothiocyanate and propidium iodide staining demonstrated that the main action of M. oleifera extract in T. gondii tachyzoites was compatible with late apoptosis. In conclusion, M. oleifera extract has anti-T. gondii activity in vitro and might be a promising substance for the development of a new anti-T. gondii drug.
刚地弓形虫是弓形体病的病原体,也是一个重要的公共卫生问题。目前,弓形体病的治疗方法是联合使用乙胺嘧啶和磺胺嘧啶,但它们在弓形体病的慢性期不起作用,而且有几种副作用。本研究评估了辣木种子水提物在体外的抗弓形体活性和潜在机制。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物细胞活力测定法确定 HeLa 细胞中辣木提取物的浓度。通过定量聚合酶链反应和甲苯胺蓝染色来评估弓形体的存在。乙胺嘧啶和磺胺嘧啶被用作药物对照。通过电子显微镜观察弓形体形态和超微结构的变化。在体外,辣木提取物在低于 50μg mL−1 的浓度下对 HeLa 细胞没有毒性作用。辣木提取物抑制弓形体的侵袭和细胞内增殖,与磺胺嘧啶+乙胺嘧啶的结果相似,并且在 30μg mL−1 的浓度下还显示细胞产生一氧化氮。电子显微镜分析表明,在处理后速殖子的结构和超微结构发生了变化。我们还观察到活性氧物质的产生增加和线粒体膜完整性的丧失。尼罗红染色试验表明脂质积累。 Annexin V-荧光素异硫氰酸酯和碘化丙啶染色表明,辣木提取物在弓形体速殖子中的主要作用与晚期细胞凋亡相吻合。总之,辣木提取物在体外具有抗弓形体活性,可能是开发新型抗弓形体药物的有前途的物质。